• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2064-2070
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• 2018

Pseudomonas tolaasii 6264 is a representative strain that causes bacterial blotch disease on the cultivated oyster mushroom, Pleurotus ostreatus. Bacteriophages are able to sterilize the pathogenic P. tolaasii strains, and therefore, they can be applied in creating disease-free mushroom cultivation farms, through a method known as "phage therapy". For successful phage therapy, the characterization of phage-resistant strains is necessary, since they are frequently induced from the original pathogenic bacteria in the presence of phages. When 10 different phages were incubated with P. tolaasii 6264, their corresponding phage-resistant strains were obtained. In this study, changes in pathogenic, genetic, and biochemical characteristics as well as the acquired phage resistance of these strains were investigated. In the phylogenetic analyses, all phage-resistant strains were identical to the original parent strain based on the sequence comparison of 16S rRNA genes. When various phage-resistant strains were examined by three different methods, pitting test, white line test, and hemolytic activity, they were divided into three groups: strains showing all positive results in three tests, two positive in the first two tests, and all negative. Nevertheless, all phage-resistant strains showed that their pathogenic activities were reduced or completely lost.

• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.83-89
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• 2001

Newcastle disease virus, CBP-1 strain isolated from Korean pheasants was passaged for 173 times by 9-day-old specific pathogenic free (SPF) embryonated eggs at $37^{\circ}C$ (parent strain) and subsequently passaged for 15 (cold attenuation (CA) -15) and 30(cold attenuation (CA) -30) times by 10-day-old of commercial broiler chicks embryonated eggs at $29^{\circ}C$, respectively, The Physical and chemical properties (sensitivity to lipid solvents, low pH and thermostability), pathogenicity (mean death time, intracerebral pathogenic index and intravenous patho-genic index), safety, booster or protective effect and characterization of temperature sensitivity were measured in cold attenuated CA-15 or 30 strain and compared to those of parent CBP-1 strain. NDV, CBP-1 CA-30 strain acquired cold attenuation and decreased infectivity at $41^{\circ}C$ compared to those of parent strain grown at $37^{\circ}C$. It lost hemagglutination activity (HA) and cell infectivity at $56^{\circ}C$ for 30, 60, and 120 Min. CA-30 strain treated with ethyl ether also lost its HA and cell infectivity. Both CA-30 and parent strains exhibited a little resistant to HA at pH 3.0 glycine HCI buffer. Intracerebral pathogenic index (ICPI) and intravenous pathogenic index (IVPI) of parent strain were 1.12 and 1.45, but decreased to 0.75 and 0.00 in CA-30 strain, respectively. The safety was evaluated by mortality in chicks inoculated with 10$^{4.0}$ EID$_{50}$ /0.1 ml. The mortalities of parent, CA-30 and commercial Bl strains were 17.5, 12.0 and 0.0%, respectively. The safety of CA-30 strain was higher than that of parent strain. The booster effects of CA-30 strain and parent strain performed in 4-week-old chicks after being vaccinated with primary commercial Bl strain.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1223-1232
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• 2017

Cordyceps militaris, a member of Ascomycota, a mushroom referred to as caterpillar Dong-chung-ha-cho, is commercially valuable because of its high content of bioactive substances, including cordycepin, and its potential for artificial cultivation. Cordycepin (3'-deoxyadenosine) is highly associated with the pharmacological effects of C. militaris. C. militaris is heterothallic in that two mating-type loci, idiomorph MAT1-1 and MAT1-2, exist discretely in two different spores. In this study, nine C. militaris strains were mated with each other to prepare newly bred strains that produced a larger amount of cordycepin than the parent strains. Nine strains of C. militaris were identified by comparing the internal transcribed spacer sequence, and a total of 12 single spores were isolated from the nine strains of C. militaris. After the MAT idiomorph was confirmed by PCR, 36 mating combinations were performed with six single spores with MAT1-1 and the others with MAT1-2. Eight mating combinations were successfully mated, producing stroma with perithecia. Cordycepin content analysis of all strains by high-performance liquid chromatography revealed that the KASP4-bred strain produced the maximum cordycepin among all strains, regardless of the medium and stroma parts. Finally, universal rice primer-PCR was performed to demonstrate that the bred strains were genetically different from the parental strains and new C. militaris strains. These results may be related to the recombination of genes during mating. The newly produced strains can be used to meet the industrial demand for cordycepin. In addition, breeding through mating suggests the possibility of producing numerous cordycepin-producing C. militaris strains.

• KSBB Journal
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• v.14 no.6
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• pp.682-687
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• 1999

Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the production of virginiamycin in Streptomyces virginiae. In order to investigate the function of VB-C as inducer in other strains, Streptomyces erythraeus was used as a test strain(parent). VB-C binding receptor gene was introduced into S. erythraeus(transformant) and the production of VBs and specific VB-C binding protein were analysed in parent and transformant. When 300ng/ml of the synthetic VB-C was added at 0, 20, 44 h cultivation of the parent and at 44 h cultivation of the transformant, the initial production times a antibiotics were shortened by more than 8 and 6 h, respectively. The transformant showed strong antibiotic activity against B. subtilis. These results suggest that the VB-C might have an ability to induce the production of secondary metabolites in S. erythraeus.

• Journal of Mushroom
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• v.12 no.3
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• pp.171-180
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• 2014

The primary objective of the present study is the characterization of the hybrids of monokaryon- monokaryon (mono-mono) crosses in mushroom breeding. We employed this technique for developing superior strains from Pleurotus species strains with 85 mono-mono intraspecific hybrids of 7 combinations between six Pleurotus ostreatus strains and one Pleurotus florida strain. In this study, the results of analysis on hybridization rate, nuclear DNA patterns, and colors and yields of fruit-bodies, are presented as follows. The crossability between mono-mono crossing ranges between 50 and 93.75%. The results of the analysis on the nuclear DNA patterns of 85 hybrid strains of mono-mono crosses share the nuclei of both parents, but their genetic similarities were predominated by either parent. The hybrid strain between P. florida and P. ostreatus showed patterns more similar to P. florida, while the hybrid strain between P. ostreatus and P. ostreatus either had patterns predominated by either parent strain. The fruiting body colors of the mono-mono crosses mostly had combined colors of both parents but showed the tendency of being more similar to that of either parent. 82% of the hybrid strain indicated similar fruiting body yields compared to both parent strains, while 0% was higher and 18% were lower than both parents. The present study was able to find out and suggest superior hybrid trains by identifying the nuclear DNA patterns of hybrids between Pleurotus species as well as the characteristics of their fruiting bodies. This study expects that the advantages of the mono-mono crossing are needs to be fully utilized in mushroom breeding and it is better to develop superior strains of Pleurotus species strains together with the mono-mono crossing.

• Korean Journal of Microbiology
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• v.36 no.4
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• pp.259-266
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• 2000

Spheroplast cell fusions were performed with Flavimonas oryzihabitans degrading aniline and Pseudomonas sp. degrading 4-chlorobiphenyl to develope the new fusant degrading aniline and 4-CBP and its characters were investigated. F. oryzihabitans was induced to antibiotic marker ($Cm^r$ by NTG treatment for the fusants selection. The results of spheroplast formation and regeneration frequencies of the strains treated with lysozyme-EDTA were 99% and 5.0~6.6%, respectively. Fusion products were treated with 40% (v/v) PEG 6000 and fusion frequency was $3.16{\times}10^{-4} $. The DNA content of fusant, F22 was approximately 2-fold compared with parents. The fusant was stable, and showed the mixed biochemical characteristics of the parent strains. F22 was similar to parent for cell growth pattern and degrading capacity on 5 mM aniline but cell growth rate of F22 was 1.5-fold higher than that of the parent on 10mM aniline. However 4-CBP degrading ability of F22 was slightly lower than that of parental strain.

• Korean Journal of Microbiology
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• v.3 no.2
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• pp.9-14
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• 1965

The enzyme-producing potentials of industrially important strains of Aspergillus spp. were studied. Irradiation of three original isolates of Aspergillus oryzae to ultra-violet rays resulted in the production of mutants which differed from the parent riboflavin and vitamin $B_{12}$ in culture media. 1. Irradition three strains of Aspergillus oryzae to ultraviolet light produced mutants and two strains of them were selected for soy sauce brewing. 2. The two strains are the physiological mutants of Aspergillus oryzae. Both were found to have superior enzyme activity to their relatives. 3. Aspergillus oryzae UV-induced mutant 172-722 and 569-713 were more powerful than others in the production of riboflavin and vitamin $B_{12}$. The enzyme activity of these strain were high and decreased only slightly even in 20 percent solution of NaCl. 4. Aspergillus oryzae UV-induced mutant 172-722 had more powerful protease producibility in wheat bran media than in modified Czapek's solution. On the contrary, Aspergillus oryzae UV-induced mutant 569-713 had more powerful producibility of saccharogenic and dextrinogenic amylase in modified Czapek's solution than in mold bran. 5. Aspergillus oryzae UV-induced mutant 172-722 formed the spore rapidly and Aspergillus oryzae UV-induced mutant 569-713 did ordinarily. 6. It is found from the results that Aspergillus oryzae UV-induced mutant 172-722 is valuable material for the manufacture of soy sauce because of its high protease activity in 20 percent solution of NaCl. Aspergillus oryzae UV-induced mutant 569-713 is suitable for soy bean mash and for fermented red pepper sauce for its high saccharogenic and dextrinogenic amylase activity in 20 percent solution of sodium chloride.

• Journal of Microbiology and Biotechnology
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• v.3 no.2
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• pp.78-85
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• 1993

In order to elucidate the mechanism which regulates the production of cyclodextrin glucanotransferase (CGTase) and to achieve overproduction of CGTase by releasing catabolite (glucose) repression, several catabolite repression resistant mutants were selected from newly screened Bacillus firmus var. alkalophilus H609, after NTG (N-methyl-N -nitro-N-nitrosoguanidine) treatment, using 2-deoxyglucose as a nonmetabolizable analog of catabolite glucose and as a selection marker. Five catabolite repression resistant mutants were selected from about 30, 000 2-deoxyglucose resistant colonies. Relative catabolite repression indices of the selected mutants were in the range of 8~80% assuming 100% for parent strain. The amount of CGTase produced by the mutant strain CR41, which was 250 units/ml, was three times larger than that produced by its parent strain. The mutation seems to have occurred in the regulatory region of CGTase gene and not in the structural region or the glucose transporting system in cell membrane. The enzymatic properties of CGTase excreted from parent and mutant strains were also compared.

• Journal of Microbiology and Biotechnology
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• v.4 no.2
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• pp.155-159
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• 1994

Two Zymomonas mobilis strains (ZM63 and ZM6307), containing both lactose and galactose operons, were constructed. $\beta$-Galactosidase and galactokinase assays indicated that both operons were expressed in both strains. The transport systems available for lactose uptake by Zymomonas mobilis were investigated using $^{14}C$-labelled lactose. After the outer membrane, which was considered to be a possible barrier to lactose uptake, was disrupted by treatment with EDTA and $Ca^{2+}$ ions, some increase in lactose uptake was observed in ZM6306 ($lac^+$) and ZM6307 ($lac^+\;gal^+$), but not in the parent, ZM6. This suggested that the outer membrane of Zymomonas mobilis acts as a barrier to lactose uptake to some degree, and also that the lactose permease is operational in Zymomonas mobilis.

• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.12-16
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• 1991

Previously RP1::Tn951 which is a derivative of RP1 containing the lactose transposon Tn951 was introduced into Z. mobilis strain ZM6l00, and RP1::Tn951 was integrated into its genome to yield ZM6306. The galactose operon was incorporated into ZM6306 to yield ZM6307 for more efficient utilization of lactose. Batch culture study has been carried out on Z. mobilis strains, ZM6306 ($lac^+$ ) and ZM6307 ($lac^+$ , $gal^+$ ), which can convert lactose directly to ethanol. Using a medium containing 80 gㆍ$1^{-1}$ glucose and 40 gㆍ$1^{-1}$ lactose, it was found that ZM6306 and ZM6307 produced maximum ethanol concentration of 40 gㆍ$1^{-1}$ and 42 gㆍ$1^{-1}$, respectively, whereas parent strain ZM6 produced 37 gㆍ$1^{-1}$.